The reaction of aromatic peptides with double helical DNA. Quantitative characterisation of a two step reaction scheme.
The binding of LysTrpLys and LysTyrLys to calf thymus DNA has been investigated by the field jump method using fluorescence detection. Two separate relaxation processes, clearly distinguished on the time scale and by opposite amplitudes, are observed for the binding of LysTrpLys to DNA with approximately 30000 base pairs. The concentration dependence of the relaxation time constants demonstrates a mechanism with a bimolecular step followed by a slow intramolecular transition with a forward rate of 6.4 X 10(3) s-1 and an equilibrium constant of 11. Measurements at various degrees of peptide binding demonstrate that the binding mechanism associated with low binding rates is restricted to a rather low number of binding sites (roughly one site in 15 base pairs). The binding of LysTyrLys to the same DNA is not associated with relaxation processes of opposite amplitudes; nevertheless two processes could be identified and assigned to a two step mechanism corresponding to that observed in the case of LysTrpLys. In the presence of sonicated DNA both peptides show a single relaxation process with characteristics similar to those observed for the slow process in the binding to high molecular DNA. The data indicate that the intramolecular step is faster for low than for high molecular DNA. these results suggest an assignment of the intramolecular step to an insertion of the aromatic residues into the DNA associated with bending of the helix. The increase in the rate of the intramolecular step with decreasing chain length of the DNA may then be explained by a higher flexibility of the double helix at lower chain lengths.
D000818:Animals; D002417:Cattle; D004247:DNA; D007700:Kinetics; D008239:Lysine; D009842:Oligopeptides; D011485:Protein Binding; D013816:Thermodynamics; D013950:Thymus Gland; D014364:Tryptophan; D014443:Tyrosine
D009842:Oligopeptides; D014443:Tyrosine; D014364:Tryptophan; D004247:DNA; D008239:Lysine